Abstract:
:Among the major obstacles to pharmacological and structural studies of integral membrane proteins (MPs) are their natural scarcity and the difficulty in overproducing them in their native form. MPs can be overexpressed in the non-native state as inclusion bodies, but inducing them to achieve their functional three-dimensional structure has proven to be a major challenge. We describe here the use of an amphipathic polymer, amphipol A8-35, as a novel environment that allows both beta-barrel and alpha-helical MPs to fold to their native state, in the absence of detergents or lipids. Amphipols, which are extremely mild surfactants, appear to favor the formation of native intramolecular protein-protein interactions over intermolecular or protein-surfactant ones. The feasibility of the approach is demonstrated using as models OmpA and FomA, two outer membrane proteins from the eubacteria Escherichia coli and Fusobacterium nucleatum, respectively, and bacteriorhodopsin, a light-driven proton pump from the plasma membrane of the archaebacterium Halobacterium salinarium.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Pocanschi CL,Dahmane T,Gohon Y,Rappaport F,Apell HJ,Kleinschmidt JH,Popot JLdoi
10.1021/bi0616706subject
Has Abstractpub_date
2006-11-28 00:00:00pages
13954-61issue
47eissn
0006-2960issn
1520-4995journal_volume
45pub_type
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